ISSN 2226-3063 e-ISSN 2227-9555 Modern Phytomorphology 12: 131–133, 2018 https://doi.org/10.5281/zenodo.1812104

RESEARCH ARTICLE

New and dangerous bacterial disease of oleander savastanoi pv. Nerii in greenhouses of Ukraine

M. M. Kluchevich 1*, P. Yа. Chumak 1, Sergey M. Vigera 2 1 Zhytomyr National Agroecological University, Staryi Blvd 7, Zhytomyr, 10008, Ukraine; * [email protected] 2 National University of Bioresources and Nature Management of Ukraine, 15 Geroyev Oborony Street, Kiyev, Ukraine; [email protected]

Received : 02.10.2018 | Accepted : 20.11.2018 | Published : 26.11.2018

Abstract

Object: Study of symptoms and etiology of oleander tuberculosis causative agents and determination of causes of their appearance in conditions of the closed ground of the greenhouses of Ukraine. Methods: Samples for research were selected in greenhouses of Botanical Garden named after acad. A. V. Fomin from different kinds and forms of oleander ( oleander L.). The microbiological and phytopathological analysis was carried out after K. I. Beltyukova. Biolog GN MicroPlate system was used to identify isolated bacterial strains. Calculations for identification in relation to genus, species and other taxonomic units are based on the similarity index. Results: On oleander , the greatest disease progress shows itself on the branches of the second year of development. Up to 5 bacterial nodes of different sizes can be formed on one branch. Galls can be formed on the leaves both on the upper (62.1%) and lower (37.9%) sides. Bacteria actively spread with water from branches and leaves, especially when spraying plants from a hose. The infection degree of different varieties of oleander was not the same. Such varieties and forms as of Nerium oleander "Album", Nerium oleander "Album Plenum" suffer from bacteriosis the most. Conclusions: For the first time in Ukraine tuberculosis Pseudomonas savastanoi pv. nerii was detected on oleander (Nerium oleander L.). The infection degree of different varieties of oleander was not the same. Nerium oleander "Album", Nerium oleander "Album Plenum", are the most severely leased, Nerium oleander "Flavescens", Nerium oleander "Lacteum" are leased to the lesser extent. The spread of the disease is favored by some methods of this group of plants growing: pruning plants, branches, and leaves bruise..

Keywords: Oleander, Bacterial node, Pseudomonas savastanoi pv. nerii

Introduction inflorescences of different colors at the ends of shoots, which are capable of long and active blossoming under The famous is the oleander (Nerium oleander L.) bright sunlight. of various forms and varieties in conditions of closed Despite the phytoncidal abilities of this plant, the ground and especially of the greenhouses of Ukraine. species is often leased by pathogens, among which the Spread of the evergreen poisonous shrub with branching bacterium Pseudomonas savastanoi pv. nerii, belonging stems is primarily caused by the presence of bright to the family , has been especially

© The Author(s) 2018. Published by Andriy Novikov, State Natural History Museum NAS of Ukraine on behalf of Modern Phytomorphology. This is an open access article under the Creative Commons BY‑NC‑ND license (http://creativecommons.org/ licenses/by‑nc‑nd/4.0/) freely available on https://phytomorphology.org/. 132 | Kluchevich et al. dangerous in recent years (Iacobellis et al. 2001). Pathogens of bacterial etiology are widespread in nature and affect many plant species in the crop. Because of high plasticity and rapid adaptation of the bacteriosis pathogens, they become resistant to many chemical (e.g. streptomycin) and biological drugs (e.g. antagonist bacteria-Pantoea agglomerans, strain P10C or Bacillus subtilis, strain BsBD 170) (Bokshan & Sadlyak 2015). That is why the determination of the causes of galls appearance on various parts of oleander, determination of bacteria species that cause them in conditions of the closed ground of the greenhouses of Ukraine is very urgent. Figure 1. Symptoms and microbiology of Pseudomonas savastanoi Materials and Method pv. nerii on Nerium oleander L.: a-parenchymal nodes on the stem; b-arenchymal nodes on the leaf area; c-primary and secondary We observed the development of Pseudomonas savastanoi parenchymal nodes; d-Pseudomonas savastanoi colonies; (original photos). pv. nerii on varieties and forms of oleander (Nerium oleander L.) in the greenhouses of Botanical Garden Bacteria actively spread with water from branches named after acad. A.V. Fomin in 2015-2017. Samples and leaves, especially when spraying plants from a hose. were taken to identify the causative agent of the disease. The spread of the disease is favored by certain features The microbiological and phytopathological analysis was of cultivation of this group of plants, in particular, carried out after K.I. Beltyukova (Beltyukova et al. 1968). because they are taken out of the greenhouses outdoors Isolation of pathogens from leased samples of oleander in summer and are back in autumn. It is also known that plants was performed on the verge of affected and the obligatory method of oleander formation is a regular healthy tissues. Small pieces of the leased tissue were pruning of plants, which also favors penetration of washed with non-sterile water for 10-15 minutes and with bacteria into open wounds, especially when disinfection sterile water- for about 5 minutes. Further, it was grated of secateurs is ignored and with use of tree-pruning to a homogeneous state in a sterile porcelain mortar and paste. Mentioned above is also due to the breaking of plated with a microbiological loop in a Petri dish on the branches and leaves, which in turn contributes to the surface of potato agar (PA). Morphology of the colonies penetration of infection. was described on the 3rd-4th day with their growth on PA. Morphology of vegetative cells and spores was studied It is found with our research, conducted in the with the help of an electron microscope (Beltyukova et al. greenhouses of Botanical Garden named after A. V. 1968). Fomin, that oleander plants are leased with four species Biology GN MicroPlate system was used to identify of arthropods: two-spotted spider mite (Tetranychus isolated bacterial strains. Calculations for bacteria urticae Koch.), oleander aphid (Aphisnerii Boyerde Fons.), identification in relation to genus, species and other oleander scale (Pseudococcus longispinus Targ.) and taxonomic units are based on the similarity index mealybug hirsute (Pseudococcus longispinus Targ.) (Schaad (Beltyukova et al. 1968). et al. 2001). Taking into account that these types of insects’ damage plants due to the presence of a piercing-sucking Results apparatus, the influence of the above-mentioned species of phytophagous pests on the spread of the causative On oleander plants, the greatest disease progress shows agent of oleander tuberculosis seems to be quite real, that itself on the branches of the second year of development requires special and profound study. (Fig. 1a and 1b). Up to 5 bacterial nodes of different sizes It should be noted that next to oleanders affected by can be formed on one branch. Galls can be formed on tuberculosis, (Olea europea L.) grows, but bacteriosis the leaves both on the upper (62.1 %) and lower (37.9 %) lesion is not yet detected on this plant species. Based on sides (Fig. 1a and 1c). Manifestations of the disease are observed on reproductive organs (inflorescence) in the idea of persistence emergence in representatives of April-June. Bacteria Pseudomonas savastanoi pv. nerii were parasitic forms of microorganisms, it should be noted isolated and identified as a result of laboratory studies that they are constantly forced to solve diverse tasks Nerium oleander "Album", Nerium oleander "Album (change peculiarities of their development). Plenum", are the most severely leased by bacteriosis, Thus, parasitic forms of microorganisms can die Nerium oleander "Flavescens", Nerium oleander "Lacteum" together with the infected organism, as the result of are leased to the lesser extent. the host’s protective factors influence. In its turn, high

Modern Phytomorphology 12, 2018 Bacterial disease of oleander Pseudomonas savastanoi | 133 virulence of the microorganism can cause the death of tuberculosis was for the first time discovered in Ukraine the infected plant before the formation of reproductive in the greenhouses of Botanical Garden named afrer A. V. individuals of the pathogen itself. That is why, pathogenic Fomin in 2015 on the purchased oleander (Neriumo leander microorganisms, on the one hand, should strive to create L.). In this connection, it can be assumed that the disease forms that are resistant to the protective factors of the was introduced together with the introductory species of infected organism, and on the other hand-to create the plant. Infection degree of different oleander varieties forms with reduced virulence (Frolov 2001). was not the same. Nerium oleander "Album", Nerium Considering the bacteriosis of Pseudomonas oleander "Album Plenum", were the most severely leased, savastanoi pv. fraxini, Pseudomonas savastanoi pv. nerii, Nerium oleander "Flavescens", Nerium oleander "Lacteum" Pseudomonas savastanoi pv. oeae from the point of view were leased to the lesser extent. of their persistence occurrence, it should be noted The spread of the disease is favored by some methods that Pseudomonas savastanoi pv. nerii differs from the of this group of plants growing: pruning plants, branches introduced bacteriosis by the ability to affect reproductive and leaves bruise. organs (inflorescence) of the host plant. This may indicate References that the co-evolution of the given parasite and the host plant is in the process of becoming. Lacobellis N.S., Maloy O.C., Murray T.D. 2001. Encyclopedia of Plant By reducing the reproductive capacity of the host Pathology. John Wiley & Sons. New York. 1: 714-715. http://ftp. plant, the parasite narrows its potential possibilities for igraph.org/Book-14/03-prof-everett-blanda-i/9788176499439- further spread. Proceeding from the idea of persistence concise-encyclopedia-of-plant-pathology-63tIwPO3NRV1.pdf emergence in parasitic forms of microorganisms, the Bokshan O.Ya., Sadlyak A.M. 2015. Promising strains of microorganisms-antagonists for biological control of bacterial revealed differences indicate the incompleteness of this fungal infections. Quarantine and Plant Protection. 193-194. process in this species. Beltyukova K.I, Matashevskaya M.S., Kulikovskaya M.D., Sidorenko Discussion S.С. 1968. Methods of studying pathogens of bacterial diseases of plants. Kiev. We should note that Pseudomonas savastanoi pv. nerii. the Schaad N.W., Jones J.B., Chun W. 2001. Laboratory Guide for disease is widely spread in Italy, Spain, France, South Identification of Plant Pathogenic Bacteria. 50: 812-813. https://onlinelibrary.wiley.com/doi/full/10.1046/j.1365- Africa, Australia, the USA, on the Black Sea coast of the 3059.2001.00635.x Caucasus: Sukhumi-Sochi and it causes tuberculosis Frolov A.F. 2001. Persistence of viruses- an evolutionary phenomenon, of plants of Nerium L. genus (Chumayevskaya 1956; preserving them as a biological species in the relationship of Krushhvili 1960; Golenko 1966; Wilson & Magie 1963). "parasite-owner" with representatives of populations of the All parts of plants are leased with tuberculosis. biosphere. International Conference "Biorezursy and viruses". Kyiv: Phytosociocenter : 58. Swellings are formed on the branches and leaves and Chumayevskaya M.A. 1956. The causative agent of tuberculosis of the leased inflorescences unusually increase in size. The oleander in the USSR. Vaskhnil. 9: 37-38. http://www.agrowebcee. disease was first described in Italy. The causative agent net/fileadmin/content/awuzbekistan/files/library/Bakterialnye_ of this disease was recognized as an independent species bolezny_rastenii..pdf and it was named Ps. tonelliana (Ғег.) Krass. (Golenko Krushhvili G.D. 1960 Oleander's tuberculosis. Protection of diseases. 1966). Based on experiments with artificial infection 10: 12. and serological studies of Ps. tonelliana isolated from Golenko M.V. 1966. Diseases of plants (the basis of the doctrine of bacteriosis of plants). Moskov. http://www.agrowebcee.net/ tuberculosis leased oleander and of P. savastanoi (E. F. fileadmin/content/aw-uzbekistan/files/library/Bakterialnye_ Smith) strains Stevens from a close serological bolezny_rastenii..pdf relationship is revealed (Wilson & Magie 1963). According Wilson E.E., Magie A.R. 1963. Physiological, serological, end to modern data, the causative agent of oleander cancer is pathological evidence that Pseudomonas tonelliana is identical considered one of the forms of the main species. with Pseudomonas sawastanoi. Phytopathology. 53: 653- 659. https://www.apsnet.org/publications/phytopathology/ Pseudomonas savastanoi (E. F. Smith) Stevens: P. backissues/Documents/1972Articles/Phyto62n03_349.PDF savastanoi pv. oeae, P. savastanoi pv. fraxini, P. savastanoi Sutic D., Dowson W.J. 1963. Microbiological characteristics of some pv. nerii, P. savastanoi pv. phaseolicola. isolates and varieties of Pseudomonas sawastanoi (Smith) Bacteria of this species are sticks 0.4-0.8 × 1.2-3.0 mc or Stevens. Phytopathology. 49: 156-160. https://onlinelibrary.wiley. 0.4-0.5 × 1.2-1.7 mc (Sutic & Dowson 1963) in size, located com/doi/pdf/10.1111/j.1439-0434.1963.tb02156.x singly, in pairs or in chains, non-sporinferious, gram- Ponyak R.I., Yakovleva L.M. 1979. Bacterial diseases of forest tree species. Kyiv. http://spbftu.ru/UserFiles/Image/izvesti/29-200. negative, the colonies are transparent on the agar, they pdf are white, round, with flat edges, they are flat and grow Kavak H., Üstün N. 2009. Oleander knot caused by Pseudomonas slowly, they form brown pigment on potato (Golenko savastanoi pv. nerii in Turkey. J. Plant. Patho. 91: 701-703. http:// 1966; Ponyak & Yakovleva 1979). www.sipav.org/main/jpp/index.php/jpp/article/view/564/352 Kudela V., Safrankova I., Krejar V., Korba J. 2005. First report Conclusion of Pseudomonas savastanoi pv. nerii on oleander in the Czech Republic. Plant Protect. Sci. 41: 33-37. https://www. We note that Pseudomonas savastanoi pv. nerii. agriculturejournals.cz/publicFiles/15529.pdf Modern Phytomorphology 12, 2018